Karsten Tedin - Academia.edu (original) (raw)

Papers by Karsten Tedin

Research paper thumbnail of A Comparison of the ATP Generating Pathways Used by S. Typhimurium to Fuel Replication within Human and Murine Macrophage and Epithelial Cell Lines

PLOS ONE, 2016

The metabolism of S. Typhimurium within infected host cells plays a fundamental role in virulence... more The metabolism of S. Typhimurium within infected host cells plays a fundamental role in virulence since it enables intracellular proliferation and dissemination and affects the innate immune response. An essential requirement for the intracellular replication of S. Typhimurium is the need to regenerate ATP. The metabolic route used to fulfil this requirement is the subject of the present study. For infection models we used human and murine epithelial and macrophage cell lines. The epithelial cell lines were mIC c12 , a transimmortalised murine colon enterocyte cell line that shows many of the characteristics of a primary epithelial cell line, and HeLa cells. The model macrophage cell lines were THP-1A human monocyte/ macrophages and RAW 264.7 murine macrophages. Using a mutational approach combined with an exometabolomic analysis, we showed that neither fermentative metabolism nor anaerobic respiration play major roles in energy generation in any of the cell lines studied. Rather, we identified overflow metabolism to acetate and lactate as the foremost route by which S. Typhimurium fulfils its energy requirements.

Research paper thumbnail of MOESM1 of ESBL-plasmid carriage in E. coli enhances in vitro bacterial competition fitness and serum resistance in some strains of pandemic sequence types without overall fitness cost

Research paper thumbnail of Identification of Natural Mutations Responsible for Altered Infection Phenotypes of Salmonella enterica Clinical Isolates by Using Cell Line Infection Screens

Applied and Environmental Microbiology, 2021

Salmonella is a foodborne pathogen affecting over 200 million people and resulting in over 200,00... more Salmonella is a foodborne pathogen affecting over 200 million people and resulting in over 200,000 fatal cases per year. Its adhesion to and invasion into intestinal epithelial cells represent one of the first and key steps in the pathogenesis of salmonellosis. Still, around 35 to 40% of bacterial genes have no experimentally validated function, and their contribution to bacterial virulence, including adhesion and invasion, remains largely unknown. Therefore, the significance of this study is in the identification of new genes or gene allelic variants previously not associated with adhesion and invasion. It is well established that blocking adhesion and/or invasion would stop or hamper bacterial infection; therefore, the new findings from this study could be used in future developments of anti- Salmonella therapy targeting genes involved in these key processes. Such treatment could be a valuable alternative, as the prevalence of antibiotic-resistant bacteria is increasing very rapidly.

Research paper thumbnail of SPI2 T3SS effectors facilitate enterocyte apical to basolateral transmigration of Salmonella-containing vacuoles in vivo

Gut Microbes, 2021

Salmonella pathogenicity island (SPI) 2 type three secretion system (T3SS)-mediated effector mole... more Salmonella pathogenicity island (SPI) 2 type three secretion system (T3SS)-mediated effector molecules facilitate bacterial survival in phagocytes but their role in the intestinal epithelium in vivo remains ill-defined. Using our neonatal murine infection model in combination with SPI2 reporter technology and RNA-Seq of sorted primary enterocytes, we demonstrate expression of SPI2 effector molecules by intraepithelial Salmonella Typhimurium (S. Typhimurium). Contrary to expectation, immunostaining revealed that infection with SPI2 T3SS-mutants resulted in significantly enlarged intraepithelial Salmonella-containing vacuoles (SCV) with altered cellular positioning, suggesting impaired apical to basolateral transmigration. Also, infection with isogenic tagged S. Typhimurium strains revealed a reduced spread of intraepithelial SPI2 T3SS mutant S. Typhimurium to systemic body sites. These results suggest that SPI2 T3SS effector molecules contribute to enterocyte apical to basolateral transmigration of the SCV during the early stage of the infection.

Research paper thumbnail of Lectin-Mediated Bacterial Modulation by the Intestinal Nematode Ascaris suum

International Journal of Molecular Sciences, 2021

Ascariasis is a global health problem for humans and animals. Adult Ascaris nematodes are long-li... more Ascariasis is a global health problem for humans and animals. Adult Ascaris nematodes are long-lived in the host intestine where they interact with host cells as well as members of the microbiota resulting in chronic infections. Nematode interactions with host cells and the microbial environment are prominently mediated by parasite-secreted proteins and peptides possessing immunomodulatory and antimicrobial activities. Previously, we discovered the C-type lectin protein AsCTL-42 in the secreted products of adult Ascaris worms. Here we tested recombinant AsCTL-42 for its ability to interact with bacterial and host cells. We found that AsCTL-42 lacks bactericidal activity but neutralized bacterial cells without killing them. Treatment of bacterial cells with AsCTL-42 reduced invasion of intestinal epithelial cells by Salmonella. Furthermore, AsCTL-42 interacted with host myeloid C-type lectin receptors. Thus, AsCTL-42 is a parasite protein involved in the triad relationship between As...

Research paper thumbnail of Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition

Scientific Reports, 2018

Toll-like receptor 5 (TLR5) is activated by bacterial flagellins and plays a crucial role in the ... more Toll-like receptor 5 (TLR5) is activated by bacterial flagellins and plays a crucial role in the first-line defence against pathogenic bacteria and in immune homeostasis, and is highly conserved in vertebrate species. However, little comparative information is available on TLR5 functionality. In this study, we compared TLR5 activation using full-length and chimeric TLR5 of various vertebrate species (human, chicken, mouse, pig, cattle). Chimeric TLR5 receptors, consisting of human transmembrane and intracellular domains, linked to extracellular domains of animal origin, were generated and expressed. The comparison of chimeric TLR5s and their full-length counterparts revealed significant functional disparities. While porcine and chicken full-length TLR5s showed a strongly reduced functionality in human cells, all chimeric receptors were functional when challenged with TLR5 ligand Salmonella FliC. Using chimeric receptors as a tool allowed for the identification of ectodomain-dependent activation potential and partially host species-specific differences in response to various enteric bacterial strains and their purified flagellins. We conclude that both the extra-and intracellular determinants of TLR5 receptors are crucial for compatibility with the species expression background and hence for proper receptor functionality. TLR5 receptors with a common intracellular domain provide a useful system to investigate bacteria-and host-specific differences in receptor activation. Toll-like receptor 5 (TLR5) is a crucial determinant of pathogen-host interaction and essential for immune homeostasis 1-4. Bacterial flagellins of diverse bacteria are the molecular stimuli that ligate and activate TLR5 in various vertebrates 5-9. TLR5 recognition of bacteria also contributes to non-infectious disease. In particular in the intestinal tract of vertebrates, TLR5 mediates various functions such as shaping the microbiota and immune balance as well as contributing to metabolic tolerance 4,10. Some bacterial species avoid TLR5 recognition by changing their flagellin protein primary sequence and by structural diversification 11-14. These evolutionary adaptations might benefit their lifestyle as chronic pathogens, environmental colonizers or symbionts. In general, the recognition of TLR5 ligands is followed by TLR5 receptor dimerization and subsequent interaction of their intracellular Toll-interleukin-1 receptor (TIR) domains with TIR domains of adaptor proteins, Myeloid Differentiation primary response protein 88 (MyD88) and TIR-domain-containing adapter-inducing interferon-β (TRIF) 15 , leading to the activation of host cell signaling pathways 16,17. The MyD88-dependent intracellular signaling cascade includes activation of mitogen-activated protein (MAP) kinases and NF-κB, leading to transcription and secretion of proinflammatory cytokines 7,18-21. Feedback modulation of the signaling cascade after initial activation also leads to the expression and activation of inhibitory molecules of the pathway, such as Toll-interacting protein (Tollip) 22 , the induction of inhibitory miRNAs 23 and to the degradation of Interleukin-1 receptor-associated kinase 1 (IRAK-1) 24 , which, in a secondary line of signaling, dampens the proinflammatory response (for review: 25).

Research paper thumbnail of Contribution of the Cpx envelope stress system to metabolism and virulence regulation in Salmonella enterica serovar Typhimurium

PLOS ONE, 2019

The Cpx-envelope stress system regulates the expression of virulence factors in many Gram-negativ... more The Cpx-envelope stress system regulates the expression of virulence factors in many Gram-negative pathogens. In Salmonella enterica serovar Typhimurium deletion of the sensor kinase CpxA but not of the response regulator CpxR results in the down regulation of the key regulator for invasion, HilA encoded by the Salmonella pathogenicity island 1 (SPI-1). Here, we provide evidence that cpxA deletion interferes with dephosphorylation of CpxR resulting in increased levels of active CpxR and consequently in misregulation of target genes. 14 potential operons were identified to be under direct control of CpxR. These include the virulence determinants ecotin, the omptin PgtE, and the SPI-2 regulator SsrB. The Tat-system and the PocR regulator that together promote anaerobic respiration of tetrathionate on 1,2-propanediol are also under direct CpxR control. Notably, 1,2-propanediol represses hilA expression. Thus, our work demonstrates for the first time the involvement of the Cpx system in a complex network mediating metabolism and virulence function.

Research paper thumbnail of The Inflammatory Response to Enterotoxigenic E. coli and Probiotic E. faecium in a Coculture Model of Porcine Intestinal Epithelial and Dendritic Cells

Mediators of Inflammation, 2018

The gut epithelium constitutes an interface between the intestinal contents and the underlying gu... more The gut epithelium constitutes an interface between the intestinal contents and the underlying gut-associated lymphoid tissue (GALT) including dendritic cells (DC). Interactions of intestinal epithelial cells (IEC) and resident DC are characterized by bidirectional crosstalk mediated by various factors, such as transforming growth factor-β (TGF-β) and thymic stromal lymphopoietin (TSLP). In the present study, we aimed (1) to model the interplay of both cell types in a porcine in vitro coculture consisting of IEC (cell line IPEC-J2) and monocyte-derived DC (MoDC) and (2) to assess whether immune responses to bacteria are altered because of the interplay between IPEC-J2 cells and MoDC. With regard to the latter, we focused on the inflammasome pathway. Here, we propose caspase-13 as a promising candidate for the noncanonical inflammasome activation in pigs. We conducted challenge experiments with enterotoxigenic Escherichia coli (ETEC) and probiotic Enterococcus faecium (E. faecium) NC...

Research paper thumbnail of {"__content__"=>"Rapid Isolation of intact -containing vacuoles using paramagnetic nanoparticles.", "i"=>{"__content__"=>"Salmonella"}}

Gut pathogens, 2018

Both typhoidal and non-typhoidal infections remain a considerable cause of morbidity and mortalit... more Both typhoidal and non-typhoidal infections remain a considerable cause of morbidity and mortality globally, and impose a major socio-economic burden worldwide. A key property of all pathogenic strains is the ability to invade host cells and reside within an intracellular, vacuolar compartment called the -containing vacuole (SCV). Although the SCV is involved in both immune-evasion and intracellular replication and spread within the host, information about the host:pathogen interactions at this interface are limited, in part due to the technical difficulties involved in purification of these vacuoles. While a number of column- or gradient-based methods have been applied, cross-contamination with other host cell organelles or rupture of the labile SCV membrane has further complicated efforts to successfully isolate SCVs. Here, we report the isolation of intact SCVs using carbon-coated, paramagnetic nanoparticles. The approach permits rapid isolation of intact SCVs from human macropha...

Research paper thumbnail of {"__content__"=>"ESBL-plasmid carriage in enhances in vitro bacterial competition fitness and serum resistance in some strains of pandemic sequence types without overall fitness cost.", "i"=>{"__content__"=>"E. coli"}}

Gut pathogens, 2018

Extended spectrum beta lactamase (ESBL)-producing extraintestinal pathogenic infections are of gl... more Extended spectrum beta lactamase (ESBL)-producing extraintestinal pathogenic infections are of global interest because of their clinical and economic impact. The ESBL resistance genes disseminate through plasmids, and are found in successful global lineages such as ST131 and ST648. The carriage of plasmids has been suggested to result in a fitness burden, but recently it was shown that ESBL-plasmids enhanced virulence in pandemic ST131 and ST648 lineages without affecting their fitness. Herein, we investigated the influence of ESBL-plasmids on bacterial competition and serum resistance, both of which are essential characteristics of ExPEC during infections. Triplets of ESBL-plasmid-carrying wildtype (WT), plasmid-cured variant (PCV) and transformant (T) of five ExPEC strains of ST131 and ST648 were used for bacterial competition experiments with colicin-producing commensal , competitive adhesion experiments and serum survival. In addition, resilience after SDS, acid, osmotic challen...

Research paper thumbnail of Altered Cytokine Expression and Barrier Properties after In Vitro Infection of Porcine Epithelial Cells with Enterotoxigenic Escherichia coli and Probiotic Enterococcus faecium

Mediators of inflammation, 2017

The aim of the present study was to elucidate the effects of the probiotic feed additive Enteroco... more The aim of the present study was to elucidate the effects of the probiotic feed additive Enterococcus faecium NCIMB 10415 (E. faecium) on porcine jejunal epithelial cells (IPEC-J2) during an in vitro challenge with enterotoxigenic Escherichia coli (ETEC). Cells were incubated with E. faecium, ETEC, or both, and the effects on barrier function and structure and intra- and intercellular signaling were determined. Coincubation with E. faecium abolished the ETEC-induced decrease in transepithelial resistance (Rt) (p ≤ 0.05). No differences were seen in the expression levels of the intercellular connecting tight junction proteins examined. However, for the first time, a reorganization of the monolayer was observed in ETEC-infected cells but not in coincubated cells. ETEC induced an increase in cytotoxicity that was prevented by coincubation (p ≤ 0.05), whereas apoptosis rates were not affected by bacterial treatment. ETEC increased the mRNA expression and release of proinflammatory cytok...

Research paper thumbnail of Salmonella Co-opts Host Cell Chaperone-mediated Autophagy for Intracellular Growth

The Journal of biological chemistry, Feb 3, 2017

Salmonella enterica are invasive intracellular pathogens that replicate within a membrane-bound c... more Salmonella enterica are invasive intracellular pathogens that replicate within a membrane-bound compartment inside infected host cells known as the Salmonella-containing vacuole. How Salmonella obtains nutrients for growth within this intracellular niche despite the apparent isolation is currently not known. Recent studies have indicated the importance of glucose and related carbon sources for tissue colonization and intracellular proliferation within host cells during Salmonella infections, although none have been found to be essential. We found that wild-type Salmonella are capable of replicating within infected host cells in the absence of both exogenous sugars and/or amino acids. Furthermore, mutants defective in glucose uptake or dependent upon peptides for growth also showed no significant loss in intracellular replication, suggesting host-derived peptides can supply both carbon units and amino acids. Here, we show that intracellular Salmonella recruit the host proteins LAMP-2...

Research paper thumbnail of Novel Immunomodulatory Flagellin-Like Protein FlaC in Campylobacter jejuni and Other Campylobacterales

mSphere, 2015

Flagellins not only are important for bacterial motility but are major bacterial proteins that ca... more Flagellins not only are important for bacterial motility but are major bacterial proteins that can modulate host responses via Toll-like receptor 5 (TLR5) or other pattern recognition receptors. Campylobacterales colonizing the intestinal tracts of different host species harbor a gene coding for an unusual flagellin, FlaC, that is not involved in motility but is secreted and possesses a chimeric amino acid sequence composed of TLR5-activating and non-TLR5-activating flagellin sequences. Campylobacter jejuni FlaC activates cells to increase in cytokine expression in chicken and human cells, promotes cross-tolerance to TLR4 ligands, and alters chicken cecal microbiota. We propose that FlaC is a secreted effector flagellin that has specifically evolved to modulate the immune response in the intestinal tract in the presence of the resident microbiota and may contribute to bacterial persistence. The results also strengthen the role of the flagellar type III apparatus as a functional secr...

Research paper thumbnail of Enterococcus faeciumNCIMB 10415 Modulates Epithelial Integrity, Heat Shock Protein, and Proinflammatory Cytokine Response in Intestinal Cells

Mediators of Inflammation, 2015

Probiotics have shown positive effects on gastrointestinal diseases; they have barrier-modulating... more Probiotics have shown positive effects on gastrointestinal diseases; they have barrier-modulating effects and change the inflammatory response towards pathogens in studiesin vitro. The aim of this investigation has been to examine the response of intestinal epithelial cells toEnterococcus faeciumNCIMB 10415 (E. faecium), a probiotic positively affecting diarrhea incidence in piglets, and two pathogenicEscherichia coli(E. coli) strains, with specific focus on the probiotic modulation of the response to the pathogenic challenge. Porcine (IPEC-J2) and human (Caco-2) intestinal cells were incubated without bacteria (control), withE. faecium, with enteropathogenic (EPEC) or enterotoxigenicE. coli(ETEC) each alone or in combination withE. faecium. The ETEC strain decreased transepithelial resistance (TER) and increased IL-8 mRNA and protein expression in both cell lines compared with control cells, an effect that could be prevented by pre- and coincubation withE. faecium. Similar effects ...

Research paper thumbnail of Probiotic Treatment Decreases the Number of CD14-Expressing Cells in Porcine Milk Which Correlates with Several Intestinal Immune Parameters in the Piglets

Frontiers in Immunology, 2015

Modulating the mucosal immune system of neonates by probiotic treatment of their mothers is a pro... more Modulating the mucosal immune system of neonates by probiotic treatment of their mothers is a promising approach which can only be investigated through the use of animal models. Here, we used sows and their piglets to investigate the impact of a bacterial treatment on the sow's milk and on the neonate piglet intestinal immune system. In previous experiments, feed supplementation of sows with the probiotic Enterococcus faecium NCIMB 10415 during pregnancy and lactation had been shown to affect intestinal microbiota and cytokine expression of the offspring during the suckling and weaning periods. We therefore investigated the composition of the milk from treated sows in comparison to samples from a control group. In treated sows, the amount of lactose increased, and the somatic cell numbers were reduced. In all milk samples, the percentage of cells expressing membranous CD14 (mCD14) was greater than the fractions of immune cells, indicating expression of mCD14 on mammary epithelial cells. However, in the milk of E. faeciumtreated sows, mCD14 + cells were reduced. Furthermore, the number of CD14 + milk cells was positively correlated with the percentages of B cells and activated T cells in the ileal MLN of the piglets. This study provides evidence for the expression of mCD14 by the porcine mammary epithelium, and suggests an immunological effect of mCD14 + milk cells on the piglets' intestinal immune system. Our study further suggests that mCD14 + mammary epithelial cell populations can be modulated by probiotic feed supplementation of the sow.

Research paper thumbnail of Influence of a probiotic Enterococcus faecium strain on development of the immune system of sows and piglets

Veterinary Immunology and Immunopathology, 2005

The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the i... more The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the intestinal colonization of pigs were determined in a feeding experiment with sows and piglets. Mucosal immunity of the developing piglets was monitored by isolation and detection of intestinal lymphocyte cell populations from the proximal jejunal epithelium and the continuous Peyers patches by the use of flow cytometry. The levels of intestinal IgA in both groups of piglets were compared, as well as total IgG in the serum of sows and piglets. Feces of the sows and intestinal contents of the piglets were taken for determination of total anaerobe and coliform bacterial counts in both probiotic and control groups. Villus length and depth of the crypts were measured in the jejunum of sacrificed piglets to monitor the development of the intestinal mucosal surface amplification. Total serum IgG of the sows appeared to be unaffected. Piglets of both groups showed similar IgG levels up to 5 weeks after birth with a slight tendency toward lower values in the probiotic group. At an age of 8 weeks the total IgG levels of the probiotic animals were significantly lower (p<0.01). No differences were observed in the populations of CD4+ and CD8+ T cells in the Peyers patches. However, the levels of cytotoxic T cells (CD8+) in the jejunal epithelium of piglets of the probiotic group were significantly reduced. The depth of the jejunal crypts and length of the villi were similar in both groups, suggesting the relative T-cell population differences were not due to alterations in the epithelial cell numbers. The total anaerobe and coliform bacterial populations were not significantly affected by the probiotic treatment, either in sows or in the piglets. However, a remarkable decline in the frequency of beta-haemolytic and O141 serovars of Escherichia coli was observed in the intestinal contents of probiotic piglets, suggesting an explanation for the reduction in cytotoxic T-cell populations.

Research paper thumbnail of Effects of Bacillus cereus var. toyoi on immune parameters of pregnant sows

Veterinary Immunology and Immunopathology, 2009

Indications for the suppression of immunological functions during pregnancy have been reported in... more Indications for the suppression of immunological functions during pregnancy have been reported in humans and cattle. Changes in the immune status during pregnancy are

Research paper thumbnail of E. coli Nissle 1917 Affects Salmonella Adhesion to Porcine Intestinal Epithelial Cells

PLoS ONE, 2011

Background: The probiotic Escherichia coli strain Nissle 1917 (EcN) has been shown to interfere i... more Background: The probiotic Escherichia coli strain Nissle 1917 (EcN) has been shown to interfere in a human in vitro model with the invasion of several bacterial pathogens into epithelial cells, but the underlying molecular mechanisms are not known. Methodology/Principal Findings: In this study, we investigated the inhibitory effects of EcN on Salmonella Typhimurium invasion of porcine intestinal epithelial cells, focusing on EcN effects on the various stages of Salmonella infection including intracellular and extracellular Salmonella growth rates, virulence gene regulation, and adhesion. We show that EcN affects the initial Salmonella invasion steps by modulating Salmonella virulence gene regulation and Salmonella SiiE-mediated adhesion, but not extra-and intracellular Salmonella growth. However, the inhibitory activity of EcN against Salmonella invasion always correlated with EcN adhesion capacities. EcN mutants defective in the expression of F1C fimbriae and flagellae were less adherent and less inhibitory toward Salmonella invasion. Another E. coli strain expressing F1C fimbriae was also adherent to IPEC-J2 cells, and was similarly inhibitory against Salmonella invasion like EcN. Conclusions: We propose that EcN affects Salmonella adhesion through secretory components. This mechanism appears to be common to many E. coli strains, with strong adherence being a prerequisite for an effective reduction of SiiE-mediated Salmonella adhesion.

Research paper thumbnail of A small non-coding RNA of the invasion gene island (SPI-1) represses outer membrane protein synthesis from the Salmonella core genome

Molecular Microbiology, 2007

The Salmonella pathogenicity island (SPI-1) encodes 35 proteins involved in assembly of a type II... more The Salmonella pathogenicity island (SPI-1) encodes 35 proteins involved in assembly of a type III secretion system (T3SS) which endows Salmonella with the ability to invade eukaryotic cells. We have discovered a novel SPI-1 gene, invR, which expresses an abundant small non-coding RNA (sRNA). The invR gene, which we identified in a global search for new Salmonella sRNA genes, is activated by the major SPI-1 transcription factor, HilD, under conditions that favour host cell invasion. The RNA chaperone, Hfq, is essential for the in vivo stability of the~80 nt InvR RNA. Hfq binds InvR with high affinity in vitro, and InvR co-immunoprecipitates with FLAG epitopetagged Hfq in Salmonella extracts. Surprisingly, deletion/overexpression of invR revealed no phenotype in SPI-1 regulation. In contrast, we find that InvR represses the synthesis of the abundant OmpD porin encoded by the Salmonella core genome. As invR is conserved in the early branching Salmonella bongori, we speculate that porin repression by InvR may have aided successful establishment of the SPI-1 T3SS after horizontal acquisition in the Salmonella lineage. This study identifies the first regulatory RNA of an enterobacterial pathogenicity island, and new roles for Hfq and HilD in SPI-1 gene expression.

Research paper thumbnail of Treponema berlinense sp. nov. and Treponema porcinum sp. nov., novel spirochaetes isolated from porcine faeces

International Journal of Systematic and Evolutionary Microbiology, 2005

Limit-dilution procedures were used to isolate seven, helically coiled bacterial strains from fae... more Limit-dilution procedures were used to isolate seven, helically coiled bacterial strains from faeces of swine that constituted two unidentified taxa. Comparative 16S rRNA gene sequence analysis showed highest similarity values with species of the genusTreponemaindicating that the isolates are members of this genus. Strain 7CPL208T, as well as five further isolates, and 14V28Tdisplayed the highest 16S rRNA gene sequence similarities withTreponema pectinovorumATCC 33768T(92·3 %) andTreponema parvumOMZ 833T(89·9 %), respectively. Polar lipid profiles distinguished 7CPL208Tand 14V28Tfrom each other as well as from related species. Based on their phenotypic and genotypic distinctiveness, strains 7CPL208Tand 14V28Tare suggested to represent two novel species of the genusTreponema, for which the namesTreponema berlinensesp. nov. andTreponema porcinumsp. nov. are proposed. The type strain forTreponema berlinenseis 7CPL208T(=ATCC BAA-909T=CIP 108244T=JCM 12341T) and forTreponema porcinum14V2...

Research paper thumbnail of A Comparison of the ATP Generating Pathways Used by S. Typhimurium to Fuel Replication within Human and Murine Macrophage and Epithelial Cell Lines

PLOS ONE, 2016

The metabolism of S. Typhimurium within infected host cells plays a fundamental role in virulence... more The metabolism of S. Typhimurium within infected host cells plays a fundamental role in virulence since it enables intracellular proliferation and dissemination and affects the innate immune response. An essential requirement for the intracellular replication of S. Typhimurium is the need to regenerate ATP. The metabolic route used to fulfil this requirement is the subject of the present study. For infection models we used human and murine epithelial and macrophage cell lines. The epithelial cell lines were mIC c12 , a transimmortalised murine colon enterocyte cell line that shows many of the characteristics of a primary epithelial cell line, and HeLa cells. The model macrophage cell lines were THP-1A human monocyte/ macrophages and RAW 264.7 murine macrophages. Using a mutational approach combined with an exometabolomic analysis, we showed that neither fermentative metabolism nor anaerobic respiration play major roles in energy generation in any of the cell lines studied. Rather, we identified overflow metabolism to acetate and lactate as the foremost route by which S. Typhimurium fulfils its energy requirements.

Research paper thumbnail of MOESM1 of ESBL-plasmid carriage in E. coli enhances in vitro bacterial competition fitness and serum resistance in some strains of pandemic sequence types without overall fitness cost

Research paper thumbnail of Identification of Natural Mutations Responsible for Altered Infection Phenotypes of Salmonella enterica Clinical Isolates by Using Cell Line Infection Screens

Applied and Environmental Microbiology, 2021

Salmonella is a foodborne pathogen affecting over 200 million people and resulting in over 200,00... more Salmonella is a foodborne pathogen affecting over 200 million people and resulting in over 200,000 fatal cases per year. Its adhesion to and invasion into intestinal epithelial cells represent one of the first and key steps in the pathogenesis of salmonellosis. Still, around 35 to 40% of bacterial genes have no experimentally validated function, and their contribution to bacterial virulence, including adhesion and invasion, remains largely unknown. Therefore, the significance of this study is in the identification of new genes or gene allelic variants previously not associated with adhesion and invasion. It is well established that blocking adhesion and/or invasion would stop or hamper bacterial infection; therefore, the new findings from this study could be used in future developments of anti- Salmonella therapy targeting genes involved in these key processes. Such treatment could be a valuable alternative, as the prevalence of antibiotic-resistant bacteria is increasing very rapidly.

Research paper thumbnail of SPI2 T3SS effectors facilitate enterocyte apical to basolateral transmigration of Salmonella-containing vacuoles in vivo

Gut Microbes, 2021

Salmonella pathogenicity island (SPI) 2 type three secretion system (T3SS)-mediated effector mole... more Salmonella pathogenicity island (SPI) 2 type three secretion system (T3SS)-mediated effector molecules facilitate bacterial survival in phagocytes but their role in the intestinal epithelium in vivo remains ill-defined. Using our neonatal murine infection model in combination with SPI2 reporter technology and RNA-Seq of sorted primary enterocytes, we demonstrate expression of SPI2 effector molecules by intraepithelial Salmonella Typhimurium (S. Typhimurium). Contrary to expectation, immunostaining revealed that infection with SPI2 T3SS-mutants resulted in significantly enlarged intraepithelial Salmonella-containing vacuoles (SCV) with altered cellular positioning, suggesting impaired apical to basolateral transmigration. Also, infection with isogenic tagged S. Typhimurium strains revealed a reduced spread of intraepithelial SPI2 T3SS mutant S. Typhimurium to systemic body sites. These results suggest that SPI2 T3SS effector molecules contribute to enterocyte apical to basolateral transmigration of the SCV during the early stage of the infection.

Research paper thumbnail of Lectin-Mediated Bacterial Modulation by the Intestinal Nematode Ascaris suum

International Journal of Molecular Sciences, 2021

Ascariasis is a global health problem for humans and animals. Adult Ascaris nematodes are long-li... more Ascariasis is a global health problem for humans and animals. Adult Ascaris nematodes are long-lived in the host intestine where they interact with host cells as well as members of the microbiota resulting in chronic infections. Nematode interactions with host cells and the microbial environment are prominently mediated by parasite-secreted proteins and peptides possessing immunomodulatory and antimicrobial activities. Previously, we discovered the C-type lectin protein AsCTL-42 in the secreted products of adult Ascaris worms. Here we tested recombinant AsCTL-42 for its ability to interact with bacterial and host cells. We found that AsCTL-42 lacks bactericidal activity but neutralized bacterial cells without killing them. Treatment of bacterial cells with AsCTL-42 reduced invasion of intestinal epithelial cells by Salmonella. Furthermore, AsCTL-42 interacted with host myeloid C-type lectin receptors. Thus, AsCTL-42 is a parasite protein involved in the triad relationship between As...

Research paper thumbnail of Functional expression of TLR5 of different vertebrate species and diversification in intestinal pathogen recognition

Scientific Reports, 2018

Toll-like receptor 5 (TLR5) is activated by bacterial flagellins and plays a crucial role in the ... more Toll-like receptor 5 (TLR5) is activated by bacterial flagellins and plays a crucial role in the first-line defence against pathogenic bacteria and in immune homeostasis, and is highly conserved in vertebrate species. However, little comparative information is available on TLR5 functionality. In this study, we compared TLR5 activation using full-length and chimeric TLR5 of various vertebrate species (human, chicken, mouse, pig, cattle). Chimeric TLR5 receptors, consisting of human transmembrane and intracellular domains, linked to extracellular domains of animal origin, were generated and expressed. The comparison of chimeric TLR5s and their full-length counterparts revealed significant functional disparities. While porcine and chicken full-length TLR5s showed a strongly reduced functionality in human cells, all chimeric receptors were functional when challenged with TLR5 ligand Salmonella FliC. Using chimeric receptors as a tool allowed for the identification of ectodomain-dependent activation potential and partially host species-specific differences in response to various enteric bacterial strains and their purified flagellins. We conclude that both the extra-and intracellular determinants of TLR5 receptors are crucial for compatibility with the species expression background and hence for proper receptor functionality. TLR5 receptors with a common intracellular domain provide a useful system to investigate bacteria-and host-specific differences in receptor activation. Toll-like receptor 5 (TLR5) is a crucial determinant of pathogen-host interaction and essential for immune homeostasis 1-4. Bacterial flagellins of diverse bacteria are the molecular stimuli that ligate and activate TLR5 in various vertebrates 5-9. TLR5 recognition of bacteria also contributes to non-infectious disease. In particular in the intestinal tract of vertebrates, TLR5 mediates various functions such as shaping the microbiota and immune balance as well as contributing to metabolic tolerance 4,10. Some bacterial species avoid TLR5 recognition by changing their flagellin protein primary sequence and by structural diversification 11-14. These evolutionary adaptations might benefit their lifestyle as chronic pathogens, environmental colonizers or symbionts. In general, the recognition of TLR5 ligands is followed by TLR5 receptor dimerization and subsequent interaction of their intracellular Toll-interleukin-1 receptor (TIR) domains with TIR domains of adaptor proteins, Myeloid Differentiation primary response protein 88 (MyD88) and TIR-domain-containing adapter-inducing interferon-β (TRIF) 15 , leading to the activation of host cell signaling pathways 16,17. The MyD88-dependent intracellular signaling cascade includes activation of mitogen-activated protein (MAP) kinases and NF-κB, leading to transcription and secretion of proinflammatory cytokines 7,18-21. Feedback modulation of the signaling cascade after initial activation also leads to the expression and activation of inhibitory molecules of the pathway, such as Toll-interacting protein (Tollip) 22 , the induction of inhibitory miRNAs 23 and to the degradation of Interleukin-1 receptor-associated kinase 1 (IRAK-1) 24 , which, in a secondary line of signaling, dampens the proinflammatory response (for review: 25).

Research paper thumbnail of Contribution of the Cpx envelope stress system to metabolism and virulence regulation in Salmonella enterica serovar Typhimurium

PLOS ONE, 2019

The Cpx-envelope stress system regulates the expression of virulence factors in many Gram-negativ... more The Cpx-envelope stress system regulates the expression of virulence factors in many Gram-negative pathogens. In Salmonella enterica serovar Typhimurium deletion of the sensor kinase CpxA but not of the response regulator CpxR results in the down regulation of the key regulator for invasion, HilA encoded by the Salmonella pathogenicity island 1 (SPI-1). Here, we provide evidence that cpxA deletion interferes with dephosphorylation of CpxR resulting in increased levels of active CpxR and consequently in misregulation of target genes. 14 potential operons were identified to be under direct control of CpxR. These include the virulence determinants ecotin, the omptin PgtE, and the SPI-2 regulator SsrB. The Tat-system and the PocR regulator that together promote anaerobic respiration of tetrathionate on 1,2-propanediol are also under direct CpxR control. Notably, 1,2-propanediol represses hilA expression. Thus, our work demonstrates for the first time the involvement of the Cpx system in a complex network mediating metabolism and virulence function.

Research paper thumbnail of The Inflammatory Response to Enterotoxigenic E. coli and Probiotic E. faecium in a Coculture Model of Porcine Intestinal Epithelial and Dendritic Cells

Mediators of Inflammation, 2018

The gut epithelium constitutes an interface between the intestinal contents and the underlying gu... more The gut epithelium constitutes an interface between the intestinal contents and the underlying gut-associated lymphoid tissue (GALT) including dendritic cells (DC). Interactions of intestinal epithelial cells (IEC) and resident DC are characterized by bidirectional crosstalk mediated by various factors, such as transforming growth factor-β (TGF-β) and thymic stromal lymphopoietin (TSLP). In the present study, we aimed (1) to model the interplay of both cell types in a porcine in vitro coculture consisting of IEC (cell line IPEC-J2) and monocyte-derived DC (MoDC) and (2) to assess whether immune responses to bacteria are altered because of the interplay between IPEC-J2 cells and MoDC. With regard to the latter, we focused on the inflammasome pathway. Here, we propose caspase-13 as a promising candidate for the noncanonical inflammasome activation in pigs. We conducted challenge experiments with enterotoxigenic Escherichia coli (ETEC) and probiotic Enterococcus faecium (E. faecium) NC...

Research paper thumbnail of {"__content__"=>"Rapid Isolation of intact -containing vacuoles using paramagnetic nanoparticles.", "i"=>{"__content__"=>"Salmonella"}}

Gut pathogens, 2018

Both typhoidal and non-typhoidal infections remain a considerable cause of morbidity and mortalit... more Both typhoidal and non-typhoidal infections remain a considerable cause of morbidity and mortality globally, and impose a major socio-economic burden worldwide. A key property of all pathogenic strains is the ability to invade host cells and reside within an intracellular, vacuolar compartment called the -containing vacuole (SCV). Although the SCV is involved in both immune-evasion and intracellular replication and spread within the host, information about the host:pathogen interactions at this interface are limited, in part due to the technical difficulties involved in purification of these vacuoles. While a number of column- or gradient-based methods have been applied, cross-contamination with other host cell organelles or rupture of the labile SCV membrane has further complicated efforts to successfully isolate SCVs. Here, we report the isolation of intact SCVs using carbon-coated, paramagnetic nanoparticles. The approach permits rapid isolation of intact SCVs from human macropha...

Research paper thumbnail of {"__content__"=>"ESBL-plasmid carriage in enhances in vitro bacterial competition fitness and serum resistance in some strains of pandemic sequence types without overall fitness cost.", "i"=>{"__content__"=>"E. coli"}}

Gut pathogens, 2018

Extended spectrum beta lactamase (ESBL)-producing extraintestinal pathogenic infections are of gl... more Extended spectrum beta lactamase (ESBL)-producing extraintestinal pathogenic infections are of global interest because of their clinical and economic impact. The ESBL resistance genes disseminate through plasmids, and are found in successful global lineages such as ST131 and ST648. The carriage of plasmids has been suggested to result in a fitness burden, but recently it was shown that ESBL-plasmids enhanced virulence in pandemic ST131 and ST648 lineages without affecting their fitness. Herein, we investigated the influence of ESBL-plasmids on bacterial competition and serum resistance, both of which are essential characteristics of ExPEC during infections. Triplets of ESBL-plasmid-carrying wildtype (WT), plasmid-cured variant (PCV) and transformant (T) of five ExPEC strains of ST131 and ST648 were used for bacterial competition experiments with colicin-producing commensal , competitive adhesion experiments and serum survival. In addition, resilience after SDS, acid, osmotic challen...

Research paper thumbnail of Altered Cytokine Expression and Barrier Properties after In Vitro Infection of Porcine Epithelial Cells with Enterotoxigenic Escherichia coli and Probiotic Enterococcus faecium

Mediators of inflammation, 2017

The aim of the present study was to elucidate the effects of the probiotic feed additive Enteroco... more The aim of the present study was to elucidate the effects of the probiotic feed additive Enterococcus faecium NCIMB 10415 (E. faecium) on porcine jejunal epithelial cells (IPEC-J2) during an in vitro challenge with enterotoxigenic Escherichia coli (ETEC). Cells were incubated with E. faecium, ETEC, or both, and the effects on barrier function and structure and intra- and intercellular signaling were determined. Coincubation with E. faecium abolished the ETEC-induced decrease in transepithelial resistance (Rt) (p ≤ 0.05). No differences were seen in the expression levels of the intercellular connecting tight junction proteins examined. However, for the first time, a reorganization of the monolayer was observed in ETEC-infected cells but not in coincubated cells. ETEC induced an increase in cytotoxicity that was prevented by coincubation (p ≤ 0.05), whereas apoptosis rates were not affected by bacterial treatment. ETEC increased the mRNA expression and release of proinflammatory cytok...

Research paper thumbnail of Salmonella Co-opts Host Cell Chaperone-mediated Autophagy for Intracellular Growth

The Journal of biological chemistry, Feb 3, 2017

Salmonella enterica are invasive intracellular pathogens that replicate within a membrane-bound c... more Salmonella enterica are invasive intracellular pathogens that replicate within a membrane-bound compartment inside infected host cells known as the Salmonella-containing vacuole. How Salmonella obtains nutrients for growth within this intracellular niche despite the apparent isolation is currently not known. Recent studies have indicated the importance of glucose and related carbon sources for tissue colonization and intracellular proliferation within host cells during Salmonella infections, although none have been found to be essential. We found that wild-type Salmonella are capable of replicating within infected host cells in the absence of both exogenous sugars and/or amino acids. Furthermore, mutants defective in glucose uptake or dependent upon peptides for growth also showed no significant loss in intracellular replication, suggesting host-derived peptides can supply both carbon units and amino acids. Here, we show that intracellular Salmonella recruit the host proteins LAMP-2...

Research paper thumbnail of Novel Immunomodulatory Flagellin-Like Protein FlaC in Campylobacter jejuni and Other Campylobacterales

mSphere, 2015

Flagellins not only are important for bacterial motility but are major bacterial proteins that ca... more Flagellins not only are important for bacterial motility but are major bacterial proteins that can modulate host responses via Toll-like receptor 5 (TLR5) or other pattern recognition receptors. Campylobacterales colonizing the intestinal tracts of different host species harbor a gene coding for an unusual flagellin, FlaC, that is not involved in motility but is secreted and possesses a chimeric amino acid sequence composed of TLR5-activating and non-TLR5-activating flagellin sequences. Campylobacter jejuni FlaC activates cells to increase in cytokine expression in chicken and human cells, promotes cross-tolerance to TLR4 ligands, and alters chicken cecal microbiota. We propose that FlaC is a secreted effector flagellin that has specifically evolved to modulate the immune response in the intestinal tract in the presence of the resident microbiota and may contribute to bacterial persistence. The results also strengthen the role of the flagellar type III apparatus as a functional secr...

Research paper thumbnail of Enterococcus faeciumNCIMB 10415 Modulates Epithelial Integrity, Heat Shock Protein, and Proinflammatory Cytokine Response in Intestinal Cells

Mediators of Inflammation, 2015

Probiotics have shown positive effects on gastrointestinal diseases; they have barrier-modulating... more Probiotics have shown positive effects on gastrointestinal diseases; they have barrier-modulating effects and change the inflammatory response towards pathogens in studiesin vitro. The aim of this investigation has been to examine the response of intestinal epithelial cells toEnterococcus faeciumNCIMB 10415 (E. faecium), a probiotic positively affecting diarrhea incidence in piglets, and two pathogenicEscherichia coli(E. coli) strains, with specific focus on the probiotic modulation of the response to the pathogenic challenge. Porcine (IPEC-J2) and human (Caco-2) intestinal cells were incubated without bacteria (control), withE. faecium, with enteropathogenic (EPEC) or enterotoxigenicE. coli(ETEC) each alone or in combination withE. faecium. The ETEC strain decreased transepithelial resistance (TER) and increased IL-8 mRNA and protein expression in both cell lines compared with control cells, an effect that could be prevented by pre- and coincubation withE. faecium. Similar effects ...

Research paper thumbnail of Probiotic Treatment Decreases the Number of CD14-Expressing Cells in Porcine Milk Which Correlates with Several Intestinal Immune Parameters in the Piglets

Frontiers in Immunology, 2015

Modulating the mucosal immune system of neonates by probiotic treatment of their mothers is a pro... more Modulating the mucosal immune system of neonates by probiotic treatment of their mothers is a promising approach which can only be investigated through the use of animal models. Here, we used sows and their piglets to investigate the impact of a bacterial treatment on the sow's milk and on the neonate piglet intestinal immune system. In previous experiments, feed supplementation of sows with the probiotic Enterococcus faecium NCIMB 10415 during pregnancy and lactation had been shown to affect intestinal microbiota and cytokine expression of the offspring during the suckling and weaning periods. We therefore investigated the composition of the milk from treated sows in comparison to samples from a control group. In treated sows, the amount of lactose increased, and the somatic cell numbers were reduced. In all milk samples, the percentage of cells expressing membranous CD14 (mCD14) was greater than the fractions of immune cells, indicating expression of mCD14 on mammary epithelial cells. However, in the milk of E. faeciumtreated sows, mCD14 + cells were reduced. Furthermore, the number of CD14 + milk cells was positively correlated with the percentages of B cells and activated T cells in the ileal MLN of the piglets. This study provides evidence for the expression of mCD14 by the porcine mammary epithelium, and suggests an immunological effect of mCD14 + milk cells on the piglets' intestinal immune system. Our study further suggests that mCD14 + mammary epithelial cell populations can be modulated by probiotic feed supplementation of the sow.

Research paper thumbnail of Influence of a probiotic Enterococcus faecium strain on development of the immune system of sows and piglets

Veterinary Immunology and Immunopathology, 2005

The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the i... more The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the intestinal colonization of pigs were determined in a feeding experiment with sows and piglets. Mucosal immunity of the developing piglets was monitored by isolation and detection of intestinal lymphocyte cell populations from the proximal jejunal epithelium and the continuous Peyers patches by the use of flow cytometry. The levels of intestinal IgA in both groups of piglets were compared, as well as total IgG in the serum of sows and piglets. Feces of the sows and intestinal contents of the piglets were taken for determination of total anaerobe and coliform bacterial counts in both probiotic and control groups. Villus length and depth of the crypts were measured in the jejunum of sacrificed piglets to monitor the development of the intestinal mucosal surface amplification. Total serum IgG of the sows appeared to be unaffected. Piglets of both groups showed similar IgG levels up to 5 weeks after birth with a slight tendency toward lower values in the probiotic group. At an age of 8 weeks the total IgG levels of the probiotic animals were significantly lower (p<0.01). No differences were observed in the populations of CD4+ and CD8+ T cells in the Peyers patches. However, the levels of cytotoxic T cells (CD8+) in the jejunal epithelium of piglets of the probiotic group were significantly reduced. The depth of the jejunal crypts and length of the villi were similar in both groups, suggesting the relative T-cell population differences were not due to alterations in the epithelial cell numbers. The total anaerobe and coliform bacterial populations were not significantly affected by the probiotic treatment, either in sows or in the piglets. However, a remarkable decline in the frequency of beta-haemolytic and O141 serovars of Escherichia coli was observed in the intestinal contents of probiotic piglets, suggesting an explanation for the reduction in cytotoxic T-cell populations.

Research paper thumbnail of Effects of Bacillus cereus var. toyoi on immune parameters of pregnant sows

Veterinary Immunology and Immunopathology, 2009

Indications for the suppression of immunological functions during pregnancy have been reported in... more Indications for the suppression of immunological functions during pregnancy have been reported in humans and cattle. Changes in the immune status during pregnancy are

Research paper thumbnail of E. coli Nissle 1917 Affects Salmonella Adhesion to Porcine Intestinal Epithelial Cells

PLoS ONE, 2011

Background: The probiotic Escherichia coli strain Nissle 1917 (EcN) has been shown to interfere i... more Background: The probiotic Escherichia coli strain Nissle 1917 (EcN) has been shown to interfere in a human in vitro model with the invasion of several bacterial pathogens into epithelial cells, but the underlying molecular mechanisms are not known. Methodology/Principal Findings: In this study, we investigated the inhibitory effects of EcN on Salmonella Typhimurium invasion of porcine intestinal epithelial cells, focusing on EcN effects on the various stages of Salmonella infection including intracellular and extracellular Salmonella growth rates, virulence gene regulation, and adhesion. We show that EcN affects the initial Salmonella invasion steps by modulating Salmonella virulence gene regulation and Salmonella SiiE-mediated adhesion, but not extra-and intracellular Salmonella growth. However, the inhibitory activity of EcN against Salmonella invasion always correlated with EcN adhesion capacities. EcN mutants defective in the expression of F1C fimbriae and flagellae were less adherent and less inhibitory toward Salmonella invasion. Another E. coli strain expressing F1C fimbriae was also adherent to IPEC-J2 cells, and was similarly inhibitory against Salmonella invasion like EcN. Conclusions: We propose that EcN affects Salmonella adhesion through secretory components. This mechanism appears to be common to many E. coli strains, with strong adherence being a prerequisite for an effective reduction of SiiE-mediated Salmonella adhesion.

Research paper thumbnail of A small non-coding RNA of the invasion gene island (SPI-1) represses outer membrane protein synthesis from the Salmonella core genome

Molecular Microbiology, 2007

The Salmonella pathogenicity island (SPI-1) encodes 35 proteins involved in assembly of a type II... more The Salmonella pathogenicity island (SPI-1) encodes 35 proteins involved in assembly of a type III secretion system (T3SS) which endows Salmonella with the ability to invade eukaryotic cells. We have discovered a novel SPI-1 gene, invR, which expresses an abundant small non-coding RNA (sRNA). The invR gene, which we identified in a global search for new Salmonella sRNA genes, is activated by the major SPI-1 transcription factor, HilD, under conditions that favour host cell invasion. The RNA chaperone, Hfq, is essential for the in vivo stability of the~80 nt InvR RNA. Hfq binds InvR with high affinity in vitro, and InvR co-immunoprecipitates with FLAG epitopetagged Hfq in Salmonella extracts. Surprisingly, deletion/overexpression of invR revealed no phenotype in SPI-1 regulation. In contrast, we find that InvR represses the synthesis of the abundant OmpD porin encoded by the Salmonella core genome. As invR is conserved in the early branching Salmonella bongori, we speculate that porin repression by InvR may have aided successful establishment of the SPI-1 T3SS after horizontal acquisition in the Salmonella lineage. This study identifies the first regulatory RNA of an enterobacterial pathogenicity island, and new roles for Hfq and HilD in SPI-1 gene expression.

Research paper thumbnail of Treponema berlinense sp. nov. and Treponema porcinum sp. nov., novel spirochaetes isolated from porcine faeces

International Journal of Systematic and Evolutionary Microbiology, 2005

Limit-dilution procedures were used to isolate seven, helically coiled bacterial strains from fae... more Limit-dilution procedures were used to isolate seven, helically coiled bacterial strains from faeces of swine that constituted two unidentified taxa. Comparative 16S rRNA gene sequence analysis showed highest similarity values with species of the genusTreponemaindicating that the isolates are members of this genus. Strain 7CPL208T, as well as five further isolates, and 14V28Tdisplayed the highest 16S rRNA gene sequence similarities withTreponema pectinovorumATCC 33768T(92·3 %) andTreponema parvumOMZ 833T(89·9 %), respectively. Polar lipid profiles distinguished 7CPL208Tand 14V28Tfrom each other as well as from related species. Based on their phenotypic and genotypic distinctiveness, strains 7CPL208Tand 14V28Tare suggested to represent two novel species of the genusTreponema, for which the namesTreponema berlinensesp. nov. andTreponema porcinumsp. nov. are proposed. The type strain forTreponema berlinenseis 7CPL208T(=ATCC BAA-909T=CIP 108244T=JCM 12341T) and forTreponema porcinum14V2...